Project Summary (Overall) Continuing funds are requested by 17 vision scientists (holding 13 eligible NEI R01 grants) and their personnel to support three resource/service cores and an administrative core in the newly merged Department of Ophthalmology, Visual and Anatomical Sciences (former Ophthalmology and Anatomy/Cell Biology Departments). The Administrative core will support the overall management of the Center Core. The research cores will enable and enhance vision research at Wayne State University and for one member, at nearby (15 minutes) Oakland University and another at Henry Ford Hospital (5 minutes away). Facilities requested are: lmaging/Histopathology (I/H), Immunology (I) and Tissue Culture/Molecular (TC/M). The I/H core in Scott Hall, will provide for confocal laser scanning, light microscopy, immunofluorescence, OCT and other cell imaging analyses (e.g., cell viability and quantification of plasma membrane potential), and train in equipment use. There also is assistance/training in management of digital images, slit lamp photography, and poster and publication production. The I core, at Kresge Eye Institute, will provide immunological assays and consultation to these vision researchers. It will initiate and stimulate innovative research projects that address emerging questions on the immunological basis of many ocular diseases. Among its functions, it will provide for isolation of peripheral blood cells for staining or viable cryopreservation, immunophenotyping, phagocytosis assays, ELISA and Western blot evaluation of cytokines/chemokines and other molecules from tissue lysates and culture supernates, image analysis for Western blots, data management and storage, and access to the C6 Accuri Flow cytometer and Cellometer instruments and to the Karmanos Flow facility. The TC/M facility, located in Scott Hall, will assist and train for preparation of specialized media, isolation, purification and characterization of ocular cells for primary culture, subculture and propagation of established cell lines, adherence assays, biofilm evaluation using crystal violet staining, genotyping of mice, cryopreservation of cells, and HEK cell culture for AAV vector packaging and purification, DNA cloning, site directed mutagenesis, and DNA transfection. Assistance and training is also available for techniques in molecular biology including PCR array and real time RT-PCR, storage of bacteria (including multi-drug resistant isolates), vectors and cDNA constructs. Each of the cores is staffed by well-trained research assistants and directors are NEI-R01 supported. The core directors and PI, as a team, prioritize NEl R01 supported studies, promote collaboration, and assist in gathering preliminary data for new NEI R01 submissions.